17beta-cyano-unsaturated steroids process and therapeutic method

ABSTRACT

17B-CYANO STEROIDS HAVING THE FORMULA   3-(O=),17-NC,13-R-ESTR-4-ENE WHERE A IS BETWEEN C9 AND   C10, AND B IS BETWEEN C11 AND C12   WHEREIN R IS ALKYL HAVING 1 TO 3 CARBON ATOMS, A IS SELECTED FROM THE GROUP CONSISTING OF TWO HYDROGENS AND A DOUBLE BOND AND B IS SELECTED FROM THE GROUP CONSISTING OF TWO HYDROGENS AND A DOUBLE BOND, WITH THE PROVISO THAT WHEN B IS A DOUBLE BOND, A IS A DOUBLE BOND, AS WELL AS THEIR PROCESS OF PREPARATION AND THERAPEUTIC COMPOSITIONS AND METHODS. THE 17-B CYANO STEROIDS POSSESS A PROGESTOMIMETIC ACTIVITY, AN ANTI-ESTROGENIC ACTIVITY AND AN EXOGENIC ANTI-ANDROGENIC ACTIVITY.

United States Patent Cffice 3,770,725 Patented Nov. 6, 1973 3,770,725 =17 53-CYANO-UNSATURATED STEROIDS, PROCESS AND THERAPEUTIC METHOD Jean-Claude Gasc, Bondy, and Robert Bucourt, Paris, France, assignors to Roussel-UCLAF, Paris, France No Drawing. Filed May 31, 1972, Ser. No. 258,324 Claims priority, appliciigiglilFrance, June 1, 1971,

Int. Cl. Cir/c 169/22 US. Cl. 260-2395 17 Claims ABSTRACT OF THE DISCLOSURE 17/3-cyano steroids having the formula THE PRIOR ART 1713 cyano 3 mesyloxy A -estratriene and 3- acetoxy 17 cyano A androstene are described respectively in Pure Appl. Chem. (1968), 16, pages 1 to and US. Pat. No. 2,326,756. These compounds, while structurally related to the above, are neither homology or isomers and do not have comparable physiological properties.

OBJECTS OF THE INVENTION An object of the present invention is the obtaining of novel unsaturated steroids having a 17fi-cyano group and interesting pharmacological properties.

Another object of the present invention is the obtaining of 17,8-cyano steroids having the formula wherein R is alkyl having 1 to 3 carbon atoms, A is selected from the group consisting of two hydrogens and a double bond and B is selected from the group consisting of two hydrogens and a double bond, with the proviso that when B is a double bond, A is a double bond.

A further object of the present invention is the development of a process for the preparation of the above 175- cyano steroids comprising reacting a 3,3-dilower alkyl ketal of a 13B-R-A -gonen-3,l7-dione with a cyclic ketalization agent selected from the group consisting of lower alkylene glycols and dioxolanes, reacting the re- I sultant 3-ketal steroid having the formula 1 oH-o CHr-O wherein R has the previously assigned value, R is hydrogen or alkyl having 1 to 5 carbon atoms, with a cyanidatron agent in a lower alkanol, dehydrating the resultant 17-cyano-17-hydroxy steroid having the formula OH I a ON I am wherein R and R have the above-assigned values by the action of an acidic dehydrating agent, reducing the A double bond of the resulting 17-cyano-A steroid having the formula R1 N an Hr-O wherein R and R have the above-assigned values, by selective hydrogenation in the presence of a hydrogenation catalyst, subjecting the resulting 3-ketal-17fi-cyano steroid having the formula wherein R and K; have the above-assigned values to acid hydrolysis and recovering said 17 3-cyano steroids.

A yet further object of the present invention is the development of therapeutic compositions and methods of therapeutic treatment utilizing the above 17B-cyan0 steroids.

These and other objects of the invention will become more apprent as the description thereof proceeds.

3 DESCRIPTION OF THE INVENTION The present invention relates to novel unsaturated steroids derivatives containing a 17/8-cyano group and to the process for preparation of these compounds.

The invention more particularly relates to steroid derivatives of the general Formula I:

can

in which the dotted lines represent possible double bonds and R represents an alkyl radical having 1 to 3 carbon atoms. More particularly the invention relates to 17,8- cyano steroids having the formula wherein R has the above-assigned values, 13fl-R-17ficyano-A -gonadien-3-ones of the Formula 1 wherein R has the above-assigned values and 13fi-R-17 3- cyano-A -gonen-3-ones of the Formula I wherein R has the above-assigned values.

The invention includes pharmaceutical compositions 75 containing at least one of the therapeutically active compounds included in the General Formula I, as well as therapeutic methods utilizing the pharmaceutical compositions.

The compounds of the invention are endowed with interesting pharmacological properties. They exhibit particularly a progestomimetic activity, an anti-estrogenic activity and an anti-androgenic activity of an exogenic character. The compounds and pharmaceutical compositions containing them may be utilized for the treatment of prostatic adenoma, of hyperandrogenia, of acne, of hirsutism and of the manifestations of hyperestrogenia. They may in addition be utilized in the treatment of sterility, dysmenorrhea, ovarial dystrophy, etc.

The compounds of the General Formula I may be utilized orally, perlingually, trancutaneously, rectally. They may be presented as injectable solutions or suspensions, packed in ampoules, multiple-dose vials, implants, tablets, coated tablets, sublingual tablets, capsules and suppositories.

The useful posology ranges between 1 and 500 mg. daily in adults, dependent upon the therapeutic indication and the method of administration. The compounds of General Formula I are administered to warm-blooded animals in amounts of from 0.01 to 10 mg./kg. per day. The pharmaceutical forms, such as: injectable solutions or suspensions, implants, tablets, coated tablets, sublingual tablets, capsules and suppositories are prepared by conventional processes.

The invention also relates to a process for the preparation of the compounds of the invention starting from 3,3- a1koxy-A -estren-17-ones described in French Pat. 1,325,398 and the higher homology in the 13,3 position, prepared by the same process. In this process, first B S-R- -cyano-A -gonen-3-ones, of Formula I are produced. This process comprises reacting a 3,3-di-lower-alkoxyl3p-R-A -gonen-17-one of the Formula VI lower alkyl 0 ketal (VII) wherein R has the above-assigned values and ketal is a cyclic ketal, preferably of the formula where R is alkyl having 1 to 5 carbon atoms, to the action of a cyanidation agent in a lower alkanol, reacting the resultant 3-ketal-1BB-R-l7g-hydroxy-17g-cyano-A gonene of Formula VIII ketal (VIII) wherein R and ketal have the above-assigned values, with an acidic dehydrating agent, subjecting the resultant 3-ketal-l3fi-R-l7-cyano-A -gonadiene of Formula IX ketal X) ketali wherein R and ketal have the above-assigned values, by the action of a strong acid, and recovering 13,8-R-17flcyano-A -gonen-3-one of the Formula 1 (I, with a double bond in the 4,5 position).

The ketalization agent used to obtain a cyclic ketal in the 3-position of the 13B-R-A -gonen-3,17-dione, VII, is preferably a lower alkylene glycol with 2 to 4 carbon atoms, such as ethylene glycol or propylene glycol. The reaction is carried out in the presence of an acidic catalyst.

The cyanidation agent used to obtain the cyanohydrins, VIII, is preferably an alkali metal cyanide, such as sodium or potassium cyanide. The cyanidation is then carried out in the presence of a lower alkanol, such as methanol, ethanol, propanol, isopropanol. The cyanidation agent may also be acetone cyanohydrin which is used in an alkaline medium.

After cyanidation a mixture of the two cyanohydrins is obtained, which can be dehydrated, as is. The mixture of the cyanohydrins may also be purified, especially by chromatography and the dehydration of one or the other of the isolated cyanohydrins or on the mixture of these cyanohydrins may be carried out.

The dehydrating agent, to whose action the cyanohydrin VIII is subjected, is preferably phosphorus oxychloride or thionyl chloride in the presence of pyridine,

The selective catalytic reduction by hydrogen of the 16 (17) double bond of the cyclic ketal in the 3-position of 3-ketal-13 6-R-17-cyano-A -gonadiene, IX, cannot be carried out in the presence of any catalyst. In fact, if the catalyst is too active, there is the risk of hydrogenating, more or less completely, the 5(10) double bond. In order to obtain a selective reaction, palladium on alumina, treated with lead acetate, is advantageously used [catalyst of the type used by Lindlar Helv. Chim. Acta 35, 446 (1952)].

The strong acid, utilized to effect the hydrolysis of the ketal X, is especially hydrochloric acid, sulfuric acid, perchloric acid, p-toluene sulfonic acid. This hydrolysis may be effected in the presence of one or several organic solvents, such as an alcohol or a hydrocarbon.

The invention also relates to a process of preparation of the 13/3-R-l7 3-cyano-A -gonadienes of Formula 1 This process comprises reacting the 3-keta1-13/8-R-17B- cyano-A -gonene of Formula X, obtained as outlined above, with an organic carboxylic acid, preferably one having 2 to 6 carbon atoms, brominating the resulting 13fl-R-l7;8-cyano-A -gonen-3-one of the Formula V wherein R has the above-assigned values, dehydrobrominating the intermediate dibromo compound and recovering the desired 13 3-R-17B-cyano-A -gonadien-3-one of Formula 1,; (I, with two double bonds).

The organic carboxylic acid, used to hydrolyze the cyclic ketal X to get, without a shift of the double bond, 13fl-R-17 3-cyano-A -gonen-3-one, V, is especially acetic acid, oxalic acid, citric acid, tartaric acid, etc. This hydrolysis is carried out in an organic solvent or in a mixture of organic solvents, such as hydrocarbons or ethers.

The brominatiou, followed by the dehydrobromination of 13/3-R-17fi-cyano-A -gonen-3-one, V, is effected by bromine in pyridine or by the pyridinium perbromide in pyridine.

The process of preparation of l3fl-R-17B-cyano-A gonatrien-3-one of Formula I is also provided by the invention. This process comprises reacting 13/8-R-17B- cyano-A -gonadien-3-one of Formula 1;; with a secondary linear or cyclic amine, hydrolyzing the resultant 3- enamine of the Formula II Rs wherein R has the above-assigned values and R and R represent two lower alkyls, a phenyl and a lower alkyl, alkylene having 4 to 5 carbon atoms, ethyloxaethylene and ethylazaethylene, in an acidic medium, subjecting the resultant 13fl-R-17fl-cyano-A -gonadien-3-one of the Formula III (III) wherein R has the above-assigned values, to the dehydrogenating action of a substituted parabenzoquinone, and recovering the desired 13/3-R-1719-cyano-A -gonadien-3-ones of Formula I The secondary amine, which is reacted with l3fi-R-l7 8- cyano-A -gonadien-3-one, I is preferably pyrrolidine. Also a di-lower-alkylamine, such as diethylamine or an alkylarylamine, preferably a lower alkylphenylamine, such as methylaniline may be used.

The reaction of 13 8-R-17fi-cyano-A -gonadien-3-one with the pyrrolidine is carried out in an organic solvent, preferably a lower alkanol, such as methanol, ethanol, propanol, isopropanol.

The acidic agent, used for the hydrolysis of the enamine is preferably formic acid, acetic acid, or tartaric acid. This hydrolysis is conveniently carried out in an organic solvent, such as ether.

The substituted parabenzoquinone used to effect the oxidation (dehydrogenation) of 1343-R-17B-cyano-A -gonadien-3-oue, 111, may particularly be selected 7 from the group consisting of 2,3-dichloro-5,6-dicyanoparabenzoquinone, 2,3 dibromo-S,G-dicyanO-parabenzoquinone, 2,3,5,6 tetrachloro-parabenzoquinone, 2,3 dicyano--chloroparabenzoquinone and 2,3-dicyano parabenzoquinone.

This oxidation reaction is carried out in inert organic solvent, such as methylene chloride, dichloroethane, benzene, toluene, dioxan, ethyl acetate, ethyl ether, etc.

The following examples are illustrative of the invention without being deemed limitative in any manner.

EXAMPLES The 3,3-alkoxy-5(10)-estren-17-ones used as starting materials are obtained by the process given in French Pat. No. 1,325,398. The higher homologs in the 13-position of this compound may be obtained by a process analogous to that of the above French patent.

Example I.--17fl-cyano-A -estren-3-one Step A: 3,3 ethylenedioxy-A -estren-17-one.3.85 gm. of pyridine hydrochloride were dissolved in 770 ml. of glycol. The solution was brought to 60-65 C. and 77 gm. of 3,3-dimethoxy-A -estren-17-one obtained by the process described in the French Pat. 1,325,398 were added. The mixture was agitated for 1 minute. Crystallization started and the agitation was maintained for 4 minutes at 60-65 C. 1 liter of iced water was added, then the mixture was poured into liter of iced water, and the agitation was continued for 1 hour.

The precipitate was vacuum filtered, washed with water and then dissolved in methylene chloride. The aqueous phase was decanted and the organic phase was dried over sodium sulfate and evaporated to dryness under vacu- The residue was dissolved in 210 ml. of isopropyl ether, about 70 ml. of the solvent was distilled off, and the mixture was chilled overnight. The precipitate was vacuum filtered and dried under vacuum at 80 C. 58 gm. of 3,3- ethylenedioxy-A -estren-17-one were obtained in the form of colorless crystals melting at 133 C. and soluble in chloroform, and insoluble in water.

I.R. spectra (chloroform).--Presence of complex C=O at 1,738 and 1.732 cm.- and of ketal. This compound is identical to that described in US. Pat. No. 2,806,030.

Step B: 3,3,-ethylenedoxy-17g-hydroxy 175 cyano- A -estrene.12.45 gm. of 3,3 ethylenedioxy A estren-l7-one and 25.6 gm. of potassium cyanide were dissolved with agitation in 625 ml. of methanol. 20.3 ml. of acetic acid were added and the mixture was agitated for 24 hours at room temperature. Thereafter, the reaction mixture was brought to a pH of 6 by the addition of acetic acid and poured into iced water. The aqueous mixture was extracted with methylene chloride. The aqueous phases were washed with water, dried over sodium sulfate, filtered and distilled to dryness under vacuum.

The residue was dissolved in 50 ml. of ethyl acetate at 60 C., filtered and chilled. The precipitate was vacuum filtered and dried at 80 C. under vacuum. 6.617 gm. of 3,3-ethylenedioxy-17.5-hydroxy-175-cyano A estrene were obtained in the form of colorless crystals melting at 196 C. and soluble in chloroform, and insoluble in water.

Analysis.C H NO molecular weight=343.45.- Calculated (percent): -N, 4.08. Found (percent): N, 4.2.

LR. spectra (chloroform).--Presence of C=N at 2,235 cm. of ketal and of OH.

Step C: 3,3 ethylenedioxy-17-cyano-A estradiene.--A mixture of 15 gm. of 3,3-ethylenedioxy-17g-hydroxy-17-cyano-A -estrene, 75 ml. of pyridine and 15 ml. of phosphorus oxychloride were agitated for 4 hours at 80 C. The reaction mixture was brought to room temperature, poured into a chilled saturated aqueous solution of sodium bicarbonate, and extracted with methylene chloride. The organic phases were washed with water,

dried over sodim sulfate, filtered and evaporated to dryness under vacuum.

The residue was subject to chromatography through silica gel with elution with a mixture (1:1) of ether and petroleum ether. After evaporation of the solvent, 11.44 gm. of the raw product were recovered. 800 mg. of this raw product were dissolved in 4 ml. of isopropyl ether at boiling and the solution was chilled. The precipitate was vacuum filtered, dried under vacuum at 70 C., and 395 mg. of 3,3-ethylenedioxy-7-cyano-A -estradiene were obtained in the form of colorless crystals melting at 129 C. and soluble in chloroform and ethanol, and insoluble in water.

Analysis.C H NO molecular weight=325.43.- Calculated (percent): N, 4.31. Found (percent): N, 4.6.

I.R. spectra (chloroform).-*Presence of conjugated CEN at 2,212 cmf of C=C at 1,591 cm. and of ketal. Absence of OH.

Step D: 3,3-ethylenedioxy 17 8-cyano-A -estrene.- 2 gm. of alumina containing 5% of palladium, treated with lead acetate (Lindlar type of catalyst. Helv. 35, 336, 1952), in suspension in 300 ml. of ethyl acetate were saturated with hydrogen. 10.2 gm. of 3,3-ethylenedioxy-17-cyano-A -estradiene were added under nitrogen, and 645 ml. of hydrogen were passed into the mixture at room temperature. The mixture was filtered and the filter was washed with methylene chloride. The filtrate obtained was evaporated to dryness under vacuum.

The residue was dissolved in 20 ml. of hot isopropylether, filtered and chilled. The crystals were vacuum filtered and dried under vacuum at 65 C. 8.64 gm. of 3,3-ethylenedioxy 1718 cyano-A -estrene were obtained in the form of colorless crystals melting at 109 C., and soluble in chloroform and insoluble in water.

Analysis.-C H NO molecular weight=327.45. Calculated (percent): N, 4.28. Found (percent): N, 4.4.

I.R. spectra (chloroform).-Presence of CEN at 2,231 cm. and of ketal.

Step E: 17 3-cyano-A -estren-3-one.-9 gm. of 3,3-ethylenedioxy-17 Scyano-A -estrene were agitated at room temperature for 1 hour with 180 ml. of a mixture containing 162 ml. of acetic acid, 5.4 ml. of hydrochloric acid and 12.6 ml. of water. The reaction mixture was poured into ice water, agitated for 30 minutes and vacuum filtered. The precipitate was Washed with water until the wash waters were neutral and dried at C. under vacuum.

The residue was purified by recrystallization from methanol, then from a methanol/methylene chloride (:20) mixture, 5.61 gm. of 17/3-cyano-A -estren-3-one were obtained in the form of colorless crystals melting at 199 C. and soluble in chloroform, ethanol and methanol, and insoluble in water. Specific rotation was (c.=0.5% in ethanol).

Analysis.C H NO; molecular weight=283.40.-- Calculated (percent): C, 80.52; H, 8.89; N, 4.94. Found (percent): C, 80.3; H, 9.0; N, 4.8.

LR. spectra (chloroform).-Presence of CEN at 2,236 cm." and A -3-one, C=O at 1,660 cm.- and C=C at 1,611 cmr UV. spectra (ethanol).-Max. at 240 nm. a: 17,500.

MNR spectra (Deutero chloroform).I-I., 357 Hz. 18 CH 61.5 Hz.

Example 2.-17fi-cyano-A -estradiene-3-one Step A: 17fl-cyano-A -estrene-3-one.A mixture of 500 mg. of 3,3-ethylenedioxy-17p-cyano-A -estrene (obtained in step D of Example 1) 25 ml. of a 66% aqueous solution of acetic acid and 10 ml. of ether were agitated during 25 hours at room temperature. The reaction mixture was then poured into iced water and extracted with methylene chloride. The organic phases were washed with an aqueous solution of sodium bicarbonate,

then with water, dried over sodium sulfate, filtered and evaporated to dryness in vacuo.

The residue was subjected to chromatography through silica gel with elution with a mixture 1:1) of ether and petroleum ether. After evaporation of the eluant, 300 mg. of 17/3-cyano-A -estren-3-one were obtained, which was used, as such, in the following step.

For analysis the product was recrystallized from a mixture (5:1) of isopropyl ether and methanol.

The compound occurred as colorless crystals melting at 124 C., and soluble in chloroform and methanol and insoluble in water.

Analysis.--C H NO; molecular weight=283.40. Calculated (percent): C, 80.52; H, 8.89; N, 4.94. Found (percent): C, 80.7; H, 9.2; N, 5.9.

LR. spectra (chloroform).Presence of C= at 1,716 cm. and of CEN at 2,235 cut- Step B: 17,8-cyano-A -estradien-3-one.11 gm. of 17fl-cyano-A -estren-3-one were dissolved in 110 ml. of pyridine. The solution was chilled to 0 to +5 C. 27.2 ml. of a methanolic solution of bromine, containing 24 gm. of Br per 100 ml. were added and the mixture was agitated for 18 hours at room temperature. The reaction mixture was then poured into 1.5 liter of iced water, containing 110 ml. of hydrochloric acid. The precipitate was vacuum filtered, washed with water and dried under vacuum at 70 C. I

3 gm. of the residue was recrystallized from a mixture (8:6) of isopropyl ether and methanol, then from methanol. 2.2 gm. of 17B-cyano-A -estradien-3-one were obtained in the form of colorless crystals melting at 189- 190 C., and soluble in chloroform and ethanol, and insoluble in water. The specific rotation is (c.=0.5% in chloroform).

Analysis.-C H NO; molecular weight=28l.38. Calculated (percent): C, 81.10; H, 8.24; N, 4.98. Found (percent): C, 81.3; H, 8.0; N, 5.2.

I.R. spectra (chloroform).-Presence of C..= N at 2,240 cmr' of C=O at 1,661 and 1,649 cm. and of C=C at 1,610 cmr U.V. spectra (ethanol):

Example 3.-17/3-cyano-A '"-estratrien-3-one Step A: 3 pyrrolidinyl-l7p-cyano-A -estratriene.-A mixture of 7.5 gm. of 17B-cyano-A -estradien- 3-one (obtained in Example 2), 30 ml. of methanol and 7.5 m1. of pyrrolidine was agitated for minutes at reflux, then chilled for 30 minutes. The precipitate was vacuum filtered and dried under vacuum. 7.4 gm. of 3-pyrrolidinyl-17fl-cyano-A -estratriene were obtained in the form of yellow crystals melting at 193 C. and soluble in chloroform and benzene, and insoluble in water.

Analysis.C H N molecular weight=334.49.-- 8.1% N (theory 8.3%).

LR. spectra (ch1oroform).--Presence of CEN at 2,231 cm.- and of conjugated system at 1,621 and 1,598 and 1,550 cmr- U.V. spectra (benzene).-Max at 357 nm. e=19,100.

Step B: l7fl-cyano-A -estradien-3-one.--A mixture of 7.4 gm. of 3-pyrrolidinyl-17;8-cyano-A estratriene, 74 ml. of water, 74 ml. of ether and 3.7 ml. of acetic acid was agitated for 2 hours. The ether was evaporated. The mixture was poured into iced water. The precipitate was vacuum filtered, washed with water and dried under vacuum at 50 C. 5.814 gm. of 17/3-cyano- A -estradien-3-one were obtained in the form of 10 cream-colored crystals melting at 144 C., and soluble in chloroform and ethanol, and insoluble in water.

IR. spectra (chloroform).-Presence of 0:0 at 1,720 cmr and of CEN at 2,230 cmr U.V. spectra (ethanol):

Step C: 17fl-cyano-A -estratrien-3-one.3.5 gm. of 17fi-cyano-A -estradien-3-one were dissolved in 70 ml. of dioxane, 5.65 gm. of dichlorodicyanobenzoquinone were added and the mixture was agitated for six hours under nitrogen. The reaction mixture was'then poured into water and extracted with methylene chloride. The organic phases were washed with an aqueous solution of sodium hydrosulfite, then with water, dried over sodium sulfate, filtered and evaporated to dryness.

The residue was dissolved in methylene chloride. The solution was passed over alumina and evaporated to dryness. The residue was dissolved in a mixture (5:1) of isopropyl ether and methanol at boiling, and chilled. The precipitate was vacuum filtered and dried. 1.624 gm. of 17B-cyano-A -estratrien-3-one were obtained in the form of colorless crystals melting at 146 C. and soluble in chloroform and ethanol, and insoluble in water. The specific rotation is [a] =-{75i2 (c. =0.6% in chloroform).

Analysis.--C H NO; molecular weight=279.37.- Calculated (percent): C, 81.68; H, 7.58; N, 5.01. Found (percent): C, 81.4; H, 7.5; N, 4.9.

LR. spectra (chloroform).-Presence of CEN at 2,237 cm.- of trienone O=O at 1,662 and 1,650 cm.- and C=C at 1,580 and 1,575 cm.

U.V. spectra (ethanol):

Max. at 237 nm. E{' =222 Max. at 337 nm. E}'% =1,100 or =30,700

Example 4.-Pharmacological studies (1) Determination of the progestomimetic activity: The progestomimetic activity was determined by the Clauberg test. According to this test, sexually immature rabbits are first sensitized by subcutaneous administration of estradiol benzoate for 5 days at a daily dose of 10 ,ug. They are next treated daily for 5 days with the medicament being studied. The animals are sacrificed on the sixth day and, on the uterus sections, the lace-like proliferation of the endometrium, characteristic for the progestomimetic action, are noted in MacPhail units.

The studied compounds utilized in solution in sesame seed oil containing 5% of benzyl alcohol, were subcutaneously administrated at difierent doses.

The obtained results are summarized in the following Table I.

TABLE I Daily Macdoses, Phail Product 7 units 17B-eyano-A -estren-3-one 500 2. 6 2,500 2. 8 17B-cyano-A '-est;radien-3-one 200 2. 3 17B-cyano-A -estratrien-S-one 1 1 1 the estradiol and the tested product. In the last case, the two steroids were injected at difierent points. The mice were sacrificed on the 4th day and their uterus was re- TABLE VI (e) Investigation of an uterotrophic activity of i7fl-cyano- ,,"-estratrienmoved and weighed.

Estradiol, in solution of sesame seed oil containing 5 Weight, of benzyl alcohol, was administered at a total dose of Lots Doses 7 g g? 0.27 with each in ection being a volume of 0.1 m1./ C 15 0 mouse. The tested products were utilized in solution in 27 2 sesame seed oil containing 5% of benzyl alcohol, and ad- Test product g3 fig mimstered 1n VaIIOLlS dosages in a volume of 0.1 m1./ 90 12.9

mouse.

The obtained results are summarized in the following Tables II to vn. TABLE VII 00 TABLE H (I) Anti-estrogenlc activity of 17fl-cyano A4 a n-estratrlen-(i-one Weight of (a) Investigation of an uterotro hic activity of 1713cyano-A -est;ren-3-o1 1 e Lots Doses uterus Contr 0 12.6 $353; i ii t 1 t di 1 1 6 1 021 85 1'0 110 puses IB- 0 p118 7 in D o 30 plus 0.27---. 26.3 (-53%) t 0 [4 D0 90171115027... 26.5 (-53%) EstradioL- 0. 27 52. 4 T t d d t 10 7.6

es e pro uc 2 3.3 These results of Tables II to VII show that the test products possess a considerable anti-estrogenic activity with reference to estradiol, and that they demonstrate no TA LE III uterotrophic effect by themselves. (b) Anti-estrcgenic activity 0f17fi-cyan0-A -estren-3-one (3) Exogenic anti-androgenic activity: The exogemc weight of anti-androgenic activity was determined with reference to Lots D0ses,-y uterus in mg. testosterone propionate in castrated male rats, by the Lerner method, described by Dorfman in Methods in Controls 0--- 7. 4 %str li t I t (11 I t i w 0 27 39 6 Hormones Research, H, page 320.

est 10 110 puses r8 0 P118 g 3&1 3 Young male rats, aged about 4 weeks, were castrated. Do 90 plus 0.27---. 30.0 (-41% The treatment started on the day after the castration and lasted 7 days. On the 8th day, the animals were sacrificed TABLE IV 3 and the following organs removed; prostate, seminal (c) Investigation oi an uterotrophic o Veslcles and leva'tor a activity of 17B-cyano- -estradien-3- The products studied and testosterone propionate were me utilized in sesame seed oil containing 5% of benzyl alco- Weight hol. The medicaments were administered separately sub- Lots Doses gig cutaneously, the products studied at the dose of 1 mg., testosterone propionate at the dose of 507 (daily dose per Controls..- 0 12. 8 rat) EstradioL- 0. 54.3 Til f 11 f h f Test product 270 12 1 e o owing groups 0 rats weret us ormed.

(a) a control group, which received the solvent;

(b) a group of rats, to which of testosterone propi- TABLE v onate was administered subcutaneously; (d) Anti-estrogenic activity of 17fl-cyano-AU-estradien-3-one a group of rats, to 1 of the Product Studied weight of was subcutaneously administered; Lots s 7 uterus in (d) a group of rats, which received 1 mg. of the product Comm] 128 50 studied subcutaneously and 507 of testosterone propi- Ech'nriinl 7.- 54.3 onate subcutaneously. Test product plus estradiol plus 0.27-.-- 49. 1

Do- 270 plus 0. 33.1 (-39%) The following Tables VIII to X summarize the results Do- 810 plus 0.27--- 31.1 (-42%) obtained.

TABLE VIII (a) 17fl-eyano-A ,-estradien-3-one Fresh Seminal levator ani, vesicles, Prostate, Lots Daily doses mg. mg. mg.

Controls 0 20. 9 10. 3 10. 4 Testosterone propionate 50v 38. 7 61. 3 90. 5 Product studied.-- 1 m g- 29. 9 9. 3 16. 0 Product studied plus testosterone propionate--- 1 mg. plus 50 32. 8 (-17%) 34. 1 (-45%) 58. 5 (-37%) TABLE IX (b) 17&cyano-A -estratiien-3-0ne Fresh levator Seminal ani, vesicles, Prostate, Lots Daily doses mg. mg. mg.

Controls. 0 20. 9 10. 3 10. 4 Testosterone propi n 50 38. 7 61. 3 90. 5 Product studied. 1 mg- 29. 3 12. 3 27. 9 Product studied plus tetsosterone propionate.- 1 mg. plus 507- 42. 2 32. 8 (-46%) 51. 9 (-42%) TABLE X (c) 17fl-cyano-A -estren-3-one Fresh levator Seminal ani, vesicules, Prostate, Lots Daily doses mg. mg. mg.

Controls. 31. 06 7. 36 12. 58 Testosterone propionate 50 44. 00 66. 82 103. 90 Product studied 1 mg-- 28. 00 6. 85 12. Product studied plus testosterone propionate 1 mg. plus 507- 38. 87 59. 05 (-12%) 83. 27 (20%) From these results, it can be noted that 17,8-cyano- A -estradien-3-one and 17B-cyano-A -estratrien-3-one exercise an important exogenic anti-androgenic activity at a dose of 1 mg. with reference to 50 of testosterone propionate and that the activity of 17B-cyano-A -estren-3- one, although Weaker, is still clear.

(4) Determination of the anti-gonadotrophic activity: The anti-gonadotrophic activity was determined in puberal rats, weighing about 200 gm. The product studied, utilized in solution in olive oil containing 5% of benzyl alcohol, were administered subcutaneously in a volume of 0.2 ml., at the ratio of 12 treatments in 14 days. On the 15th day, the rats were sacrificed by carotid bleeding, and the seminal vesicles, the prostate, the testicles and the suprarenals were removed and weighed.

The obtained results are summarized in the following Table XI.

TABLE XI Daily Testi- Seminal Supradoses, cles, vesicles, Prostate, renals, Lots mg. mg. mg. mg. mg.

Controls 0 2,838 529.58 320.18 40.28 17fl-eyano-N-estren-3-ona. 2 2, 670 40.98 o 2 990 864.5 451.6 46.4 17B-cyano-A estradien- 2 gggg 2 i 98%.? i /ii} 17B A4 9 u t 0 2,990 864.5 451. 6 46.4

'CYBl'lO' I -es Y8.- trien-3-one 2 2,s9o{ jag; 43.7 40

These results demonstrate that the 3 products studied possessed a certain anti-gonadotrophic activity at a daily dose of 2 mg.

The preceding specific embodiments are illustrative of the practice of the invention. It is to be understood, however, that other expedients known to those skilled in the art or disclosed herein, may be employed without departing from the spirit of the invention or the scope of the appended claims.

We claim:

1. l7 3-cyano steroids having the formula UCN l l wherein R is alkyl having 1 to 3 carbon atoms, A is selected from the group consisting of two hydrogens and a double bond and B is selected from the group consisting of two hydrogens and a double bond, with the proviso that when B is a double bond, A is a double bond.

2. The compound of claim 1 wherein A and B are both two hydrogens.

3. The compound of claim 2 being 17ficyano-A -estrcn- 3-one.

4. The compound of claim 1 wherein A is a double bond and B is two hydrogens.

5. The compound of claim 4 being l7fl-cyano-A estradien-S-one.

6. The compound of claim 1 wherein A and B are both double bonds.

Hz-O

wherein R has the previously assigned value, R is hydrogen or alkyl having 1 to 5 carbon atoms, with a cyanidation agent selected from the group consisting of (1) and alkali metal cyanide in a lower alkanol and (2) acetone cyanohydrin in an alkaline medium, dehydrating the resultant 17-cyano-l7-hydroxy steroid having the formula R I a: i ON R1 err-o CH -O wherein R and R have the above-assigned values by the action of an acidic dehydrating agent selected from the group consisting of phosphorus oxychloride and thionyl chloride, in the presence of pyridine, reducing the A -double bond of the resulting l7-cyano-A steroid having the formula R1 ta.

Ill CN CHz-O wherein R and R have the above-assigned values, by selective hydrogenation in the presence of palladium on alumina, treated with lead acetate, as a Lindlar type hydrogenation catalyst, subjecting the resulting 3-ketal-17flcyano steroid having the formula CH:O

wherein R and R have the above-assigned values to acid hydrolysis and recovering said 17 3-cyano steroids.

9. The process of claim 8 wherein said 3-ketal-17B- cyano steroid is hydrolyzed with a strong acid and a 17 8- cyano steroid where A and B are both two hydrogens is recovered.

10. The process of claim 8 wherein said 3-ketal-l7 3- cyano steroid is hydrolyzed with an organic carboxylic acid having 2 to 6 carbon atoms, the resultant 17B-cyano- M -steroid having the formula wherein R has the prior assigned values is subjected to a bromination, then dehydrobromination by means selected from the group consisting of bromine in pyridine and pyridinium perbrornide in pyridine, and a 17fl-cyano steroid where A is a double bond and B is two hydrogens is recovered.

11. The process of claim 8 wherein said 3-ketal-17B- cyano steroid is hydrolyzed with an organic carboxylic acid having 2 to 6 carbon atoms, the resultants H S-cyano- A -steroid having the formula ON OW wherein R has the prior assigned values is subjected to a bromination, then dehydrobromination by means selected from the group consisting of bromine in pyridine and pyridinium perbromide in pyridine, the resultant 17- cyano steroid where A is a double bond and B is two hydrogens is reacted with a secondary amine selected from the group consisting of di-lower alkylamines, lower alkyl-phenylamines pyrrolidine, pyrrole, morpholine and piperazine, the resultant 3-enamine having the formula R CN 3 1 H 1 iHz-O 16 wherein R is alkyl having 1 to 3 carbon atoms and R is hydrogen or alkyl having 1 to 5 carbon atoms.

13. l7-cyano-A steroids having the formula bH-o I 112-0 wherein R is alkyl having 1 to 3 carbon atoms and R is hydrogen or alkyl having 1 to 5 carbon atoms.

14. 3-ketal-17fi-cyano steroids having the formula (BE-O lHr-O wherein R is alkyl having 1 to 3 carbon atoms and R is hydrogen or alkyl having 1 to 5 carbon atoms.

15. 17fi-cyano-A steroids having the formula wherein R is alkyl having 1 to 3 carbon atoms.

16. 3-enamines having the formula LON I wherein R is alkyl having 1 to 3 carbon atoms and R and R represent members selected from the group consisting of two lower alkyls, a phenyl and a lower alkyl, alkylene having 4 to 5 carbon atoms, ethyloxaethylene and ethylazaethylene.

17. 17B-cyano-A steroids having the formula R l @N M wherein R is alkyl having 1 to 3 carbon atoms.

References Cited UNITED STATES PATENTS 2,326,756 8/1943 Butenandt et a1. 260-3975 3,301,878 1/1967 Joly et a1. 260-3973 3,383,385 5/1968 Bucourt et al. 260-23957 HENRY A. FRENCH, Primary Examiner US. Cl. X.R. 

